Analysis of Structure and Function of Ladybird Leg and Subsequent Design and Fabrication of a Simplified Leg Structure for Robotic Applications.

Many insects are able to walk vertically or upside down on both hard and soft surfaces. In beetles such as the ladybird (Coccinella septempunctata), intermolecular forces between tarsal setae on the footpads of the insects make this movement possible. In prior work, adhesion structures made from polydimethylsiloxane (PDMS) that mimic the action of the tarsal setae have been developed. It is proposed that these adhesion structures could be attached to a simplified version of the leg of a ladybird and used in practical applications. For example, the leg structures could potentially be employed in small surveillance drones to enable attachment to surfaces during flights, in order to preserve battery power. Alternatively, the structures could be used in small robotic devices to enable walking on steeply inclined surfaces. In this program of work, the morphology and movement of the leg of a ladybird were closely studied using a 3D X-ray microscope and a high-speed microscope. The positions of the tendons that facilitated movement were identified. From this knowledge, a simplified leg structure using pin-joints was designed and then fabricated using 3-D printing. The PDMS adhesion structures were then attached to the leg structure. The tendons in the actual insect leg were replicated using thread. Typical detachment forces of about 4 N indicated that the simplified leg structure was, in principle, more than capable of supporting the weight of a small device and then detach successfully. Attachment/detachment movement operations were performed using a linear actuator and controlled remotely. Therefore, proof of concept has been demonstrated for the use of such a simplified ladybird leg structure for the attachment/detachment of small robotic devices to horizontal, inclined, or vertical surfaces.

Evidence for intermolecular forces involved in ladybird beetle tarsal setae adhesion.

Why can beetles such as the ladybird beetle Coccinella septempunctata walk vertically or upside-down on a smooth glass plane? Intermolecular and/or capillary forces mediated by a secretion fluid on the hairy footpads have commonly been considered the predominant adhesion mechanism. However, the main contribution of physical phenomena to the resulting overall adhesive force has yet to be experimentally proved, because it is difficult to quantitatively analyse the pad secretion which directly affects the adhesion mechanism. We observed beetle secretion fluid by using inverted optical microscopy and cryo-scanning electron microscopy, which showed the fluid secretion layer and revealed that the contact fluid layer between the footpad and substrate was less than 10-20 nm thick, thus indicating the possibility of contribution of intermolecular forces. If intermolecular force is the main physical phenomenon of adhesion, the force will be proportional to the work of adhesion, which can be described by the sum of the square roots of dispersive and polar parts of surface free energy. We measured adhesion forces of ladybird beetle footpads to flat, smooth substrates with known surface free energies. The adhesive force was proportional to the square-root of the dispersive component of the substrate surface free energy and was not affected by the polar component. Therefore, intermolecular forces are the main adhesive component of the overall adhesion force of the ladybird beetle. The footpads adhere more strongly to surfaces with higher dispersive components, such as wax-covered plant leaves found in the natural habitat of ladybird beetles. Based on the present findings, we assume ladybird beetles have developed this improved performance as an adaptation to the variety of plant species in its habitat.

Crucial role of framework with cytoskeletal actin filaments for shaping microstructure of footpad setae in the ladybird beetle, Harmonia axyridis.

Insects that can walk on smooth surfaces have specialized structures, footpads, on their legs. Footpads play an important role in adhesion to the substrate surface. Although the morphology and function of footpads have been studied, the mechanism of their formation is still elusive. In the ladybird beetle (Harmonia axyridis), hairy footpads are present on the first and second tarsal segments of the legs. The footpads are covered with hundreds of hairs, i.e. setae, whose tips consist of four types: pointed, lanceolate, spatular, and discoidal. We examined the formation of the footpad during the pupal stage using immuno-staining and fluorescent-conjugated phalloidin staining. We found that a seta was composed of a shaft and a socket and some setae were accompanied by a neuron. By the mid-pupal stages, the shaft cells elongated to form a setal structure. Cytoskeletal actin bundles ramified to create a framework for the setal tip structure of the cells. We examined the effects of the application of cytochalasin D, which inhibits actin polymerization, on the formation of footpad setal structures. The results showed that the setal tips were deformed by the inhibition of actin polymerization. Our observations reveal that cytoskeletal actin filaments are involved in shaping the setae.

Framework with cytoskeletal actin filaments forming insect footpad hairs inspires biomimetic adhesive device design.

Footpads allow insects to walk on smooth surfaces. Specifically, liquid secretions on the footpad mediate adhesiveness through Van der Waals, Coulomb, and attractive capillary forces. Although the morphology and function of the footpad are well defined, the mechanism underlying their formation remains elusive. Here, we demonstrate that footpad hair in Drosophila is formed by the elongation of the hair cells and assembly of actin filaments. Knockdown of Actin5C caused a malformation of the hair structure, resulting in reduced ability to adhere to smooth substrates. We determined that functional footpads are created when hair cells form effective frameworks with actin filament bundles, thereby shaping the hair tip and facilitating cuticular deposition. We adapted this mechanism of microstructure formation to design a new artificial adhesive device⁠-a spatula-like fiber-framed adhesive device supported by nylon fibers with a gel material at the tip. This simple self-assembly mechanism facilitates the energy-efficient production of low-cost adhesion devices.

Hierarchical Structure of the Cocos nucifera (Coconut) Endocarp: Functional Morphology and its Influence on Fracture Toughness.

In recent years, the biomimetic potential of lignified or partially lignified fruit pericarps has moved into focus. For the transfer of functional principles into biomimetic applications, a profound understanding of the structural composition of the role models is important. The aim of this study was to qualitatively analyze and visualize the functional morphology of the coconut endocarp on several hierarchical levels, and to use these findings for a more precise evaluation of the toughening mechanisms in the endocarp. Eight hierarchical levels of the ripe coconut fruit were identified using different imaging techniques, including light and scanning electron microscopy as well as micro-computer-tomography. These range from the organ level of the fruit (H0) to the molecular composition (H7) of the endocarp components. A special focus was laid on the hierarchical levels of the endocarp (H3-H6). This investigation confirmed that all hierarchical levels influence the crack development in different ways and thus contribute to the pronounced fracture toughness of the coconut endocarp. By providing relevant morphological parameters at each hierarchical level with the associated toughening mechanisms, this lays the basis for transferring those properties into biomimetic technical applications.

Substrate-borne vibrations induce behavioral responses in the leaf-dwelling cerambycid, Paraglenea fortunei.

Many insects utilize substrate-borne vibrations as a source of information for recognizing mates or predators. Among various substrates, plant leaves are commonly used for transmitting and receiving vibrational information. However, little is known about the utilization of vibrations by leaf-dwelling insects, especially coleopteran beetles. We conducted two experiments to examine the response of the leaf-dwelling cerambycid beetle, Paraglenea fortunei, to substrate-borne vibrations. We recorded and analyzed vibrations of host plant leaves from four different sources: wind (0.5 m/s), a beetle during landing, a walking beetle, and a beetle walking in the wind (0.5 m/s). We then measured the behavioral thresholds, the lowest amplitudes that induce behavioral responses, from beetles walking and resting on horizontal and vertical substrates with pulsed vibrations ranging from 20 Hz to 1 kHz. The vibrational characteristics of biotic and abiotic stimuli clearly differed. Beetle-generated vibrations (landing, walking, and walking in the wind) were broadly high in the low-frequency components above ∼30 Hz, while wind-generated vibrations showed a dominant peak at ∼30 Hz and a steep decrease thereafter. Among four situations, beetles walking on horizontal substrates showed lowest thresholds to vibrations of 75-500 Hz, which are characteristic of beetle-generated vibrations. Given that P. fortunei beetles are found on horizontal leaf surfaces of the host plant, vibrations transmitted though horizontal substrates may induce a strong freeze response in walking beetles to detect conspecifics or heterospecifics. Our findings provide evidence that leaf-dwelling beetles can discriminate among biotic and abiotic factors via differences in vibrational characteristics.

Underwater locomotion in a terrestrial beetle: combination of surface de-wetting and capillary forces.

For the first time, we report the remarkable ability of the terrestrial leaf beetle Gastrophysa viridula to walk on solid substrates under water. These beetles have adhesive setae on their feet that produce a secretory fluid having a crucial role in adhesion on land. In air, adhesion is produced by capillary forces between the fluid-covered setae and the substrate. In general, capillary forces do not contribute to adhesion under water. However, our observations showed that these beetles may use air bubbles trapped between their adhesive setae to walk on flooded, inclined substrata or even under water. Beetle adhesion to hydrophilic surfaces under water was lower than that in air, whereas adhesion to hydrophobic surfaces under water was comparable to that in air. Oil-covered hairy pads had a pinning effect, retaining the air bubbles on their feet. Bubbles in contact with the hydrophobic substrate de-wetted the substrate and produced capillary adhesion. Additional capillary forces are generated by the pad's liquid bridges between the foot and the substrate. Inspired by this idea, we designed an artificial silicone polymer structure with underwater adhesive properties.

Friction force reduction triggers feet grooming behaviour in beetles.

In insects, cleaning (grooming) of tarsal attachment devices is essential for maintaining their adhesive ability, necessary for walking on a complex terrain of plant surfaces. How insects obtain information on the degree of contamination of their feet has remained, until recently, unclear. We carried out friction force measurements on walking beetles Gastrophysa viridula (Coleoptera, Chrysomelidae) and counted grooming occurrence on stiff polymer substrata with different degrees of nanoroughness (root mean square: 28-288 nm). Since nanoscopically, rough surfaces strongly reduced friction and adhesion without contaminating feet, we were able to demonstrate, for the first time to our knowledge, that friction force between tarsal attachment pads and the substrate provides an insect with information on the degree of contamination of its attachment structures. We have shown that foot grooming occurrence correlates not only with the degree of contamination but also with the decrease of friction force. This result indicates that insects obtain information about the degree of contamination, not statically but rather dynamically and, presumably, use mechanoreceptors monitoring either tensile/compressive forces in the cuticle or tensile forces between leg segments.

Waterproof and translucent wings at the same time: problems and solutions in butterflies.

Although the colour of butterflies attracts the most attention, the waterproofing properties of their wings are also extremely interesting. Most butterfly wings are considered "super-hydrophobic" because the contact angle (CA) with a water drop exceeds 150 degrees. Usually, butterfly wings are covered with strongly overlapping scales; however, in the case of transparent or translucent wings, scale cover is reduced; thus, the hydrophobicity could be affected. Here, we present a comparative analysis of wing hydrophobicity and its dependence on morphology for two species with translucent wings Parantica sita (Nymphalidae) and Parnassius glacialis (Papilionidae). These species have very different life histories: P. sita lives for up to 6 months as an adult and migrates over long distance, whereas P. glacialis lives for less than 1 month and does not migrate. We measured the water CA and analysed wing morphology with scanning electron microscopy and atomic force microscopy. P. sita has super-hydrophobic wing surfaces, with CA > 160 degrees, whereas P. glacialis did not (CA = 100-135 degrees). Specialised scales were found on the translucent portions of P. sita wings. These scales were ovoid and much thinner than common scales, erect at about 30 degrees, and leaving up to 80% of the wing surface uncovered. The underlying bare wing surface had a remarkable pattern of ridges and knobs. P. glacialis also had over 80% of the wing surface uncovered, but the scales were either setae-like or spade-like. The bare surface of the wing had an irregular wavy smooth pattern. We suggest a mode of action that allows this super-hydrophobic effect with an incompletely covered wing surface. The scales bend, but do not collapse, under the pressure of a water droplet, and the elastic recovery of the structure at the borders of the droplet allows a high apparent CA. Thus, P. sita can be translucent without losing its waterproof properties. This characteristic is likely necessary for the long life and migration of this species. This is the first study of some of the effects on the hydrophobicity of translucency through scales' cover reduction in butterfly wings and on the morphology associated with improved waterproofing.

Influence of surface roughness on gecko adhesion.

In this study we show the influence of surface roughness on gecko adhesion on both the nano- and macroscales. We present experimental data for the force necessary to pull off single spatulae from hard rough substrates and also detail observations on living geckos clinging to various surfaces. Both experiments consistently show that the effective adhesion shows a minimum for a root mean square roughness ranging from 100 to 300nm.